Curcumol Combined With 5-Fluorouracil In Regulation Of The Proliferation And Apoptosis Of Human Gastric Cancer SGC-7901 Cells

Objectives:To initially investigate the mechanisms of the effects of curcumol and 5-fluorouracil on the development of human gastric cancer SGC-7901 cells.Subject:Human gastric cancer SGC-7901 cellsMethods:The curcumol was dissolved in dehydrate alcohol, making the initial concentration of curcumol 5mg/ml, as it is a mother liquor. First, the experimental groups of 1、2、3、4、5 were set up as curcumol groups and concentrations were set as follows:0μg/ml、8μg/ml、16μg/ml、 32μg/ml、64μg/ml. Group 1 is a control group which contained 1% dehydrate alcohol, as a dissolvent compare. Second, the experimental groups of 1、2、3、4 were set up as curcumol combined with 5-fluorouracil groups and concentrations were set as follows:0μg/ml control group、32μg/ml curcumol group、5μ g/ml 5-fluorouracil group、curcumol(32μg/ml) combined with 5-fluorouracil(5μg/ml) group. After cells were seeded for 48h, the Trypan blue method was used to analyze the cytoactive of SGC-7901. After cells were treated with curcumol and 5-fluorouracil at varying concentrations for various hours, MTT assay was performed to study the inhibitory rate of SGC-7901 cells proliferation. After cells were treated with curcumol and 5-fluorouracil at varying concentrations for 48 hours, the cell apoptosis rate were detected by FCM, the expression of bax、bcl-2 and caspase-3 were detected by Western Blot and the enzyme activity of caspase-3 were detected by the activity detection kit of caspase-3.Results:1. The Trypan blue dyeing showed that the viable rate of SGC-7901 were more than 90%, that fitted the requirement of experiment. After cells were treated with curcumol and 5-fluorouracil at varying concentrations for various hours, the proliferation inhibition rates of cells increased significantly when compared with the control group (P<0.05). The higher the concentration of curcumol, the higher the inhibition rate of cell proliferation. Wherein, the inhibition rates of cells which were treated for 48 hours are the most obvious. The proliferation inhibition rate of curcumol combined with 5-fluorouracil group was significantly higher than monotherapy groups (P<0.05).2. After cells were treated for 48h and detected by FCM assay, it showed that the apoptosis rate of control group was about 2.37%, curcumol group 13.43%,5-fluorouracil group 10.59% and curcumol combined with 5-fluorouracil group 24.11%. Curcumol could obviously promote the apoptosis induced by 5-fluorouracil (P<0.05).3. Western Blot confirmed that curcumol and 5-fluorouracil could significantly promote the expression of bax and inhibit the expression of bcl-2 after cells were treated for 48h. Wherein, the effect of curcumol combined with 5-fluorouracil group was more obvious than monotherapy groups (P<0.05).4. Western Blot and activity detection kit of caspase-3 confirmed that curcumol and 5-fluorouracil could significantly promote the expression and enzyme activity of caspase-3 after cells were treated for 48h. Wherein, the effect of curcumol combined with 5-fluorouracil group was more obvious than monotherapy groups(P<0.05).Conclusion:1. Curcumol can significantly inhibit proliferation and induce apoptosis of SGC-7901 cells. The effect is more obvious after cells are treated combined with 5-fluorouracil. It suggests that curcumol could significantly strengthen the sensitivity of tumor cells to chemotherapeutic drugs.2. Curcumol and 5-fluorouracil can activate caspase-3 by regulating the proportion of apoptotic proteins bax/bcl-2, leading to apoptosis. The effect is more obvious by combination therapy. It suggests that curcumol promoted apoptosis induced by 5-fluorouracil possibly through the endogenous apoptotic pathway mediated by mitochondria.